Download 12 December Aula NIOB/CBS

Dear Friends, We herewith kindly invite you for the scientific meeting of the Netherlands
Society for Medical Mycology, to be held on 12 December, 2007 in the auditorium of the
Centraalbureau voor Schimmelcultures, Uppsalalaan 8, Utrecht, The Netherlands.
Participation is free of charge. Coffee and tea will be available at no costs.
PROGRAMME:
Chairman: Sybren de Hoog
09.30-09.45
Kathrin Tintelnot, Andrea Haas, Barbara Graf and M. Seibold (Robert Koch Institut,
Berlin, Germany): Scedosporium prolificans infections in Germany.
09.45-10.00
Hamid Badali1,2, Bert Gerrits van den Ende1, Derlene Attili3 and Sybren de Hoog1
(1Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands; 2Department of
Medical Mycology and Parasitology, School of Medicine, Mazandaran University of
Medical Sciences, Sari, Iran; 3Universidade Estadual Paulista, UNESP Rio Claro, SP,
Brazil): New species of Cladophialophora.
10.00-10.15
E. Snelders, H.A.L. van der Lee, J. Kuijpers, A.J.M.M. Rijs, W.J.G. Melchers and P.E.
Verweij (University Medical Center St. Radboud, Nijmegen, The Netherlands):
Molecular characterisation of azole-resistant Aspergillus fumigatus isolates.
10.15-10.30
Ignace Surmont (Heilig Hart Ziekenhuis, Roeselaere, Belgium): Ernstige locale
Aspergillus infectie bij hematologische patient.
10.30-10.45
Wendy W.J. van de Sande1, Ahmed Fahal2, Henri Verbrugh1 and Alex van Belkum1
(1Department of Bacteriology, University of Rotterdam, The Netherlands; 2Mycetoma
Research Center, University of Khartoum, Sudan): Polymorphisms in genes involved in
innate immunity predispose towards myetoma susceptibility.
10.45-11.00
Dermatophytes: To be announced.
11.00 – 11.30 BREAK
Chairman: Jacques Meis
11.30-11.45
Louis Chai1, David Denning2 and Peter Warn2 (1University Medical Center St. Radboud,
Nijmegen, The Netherlands; School of Medicine, University of Manchester, U.K.):
Candida tropicalis in human disease.
11.45-12.00
Hanneke de Valk et al. (Department of Medical Microbiology and Infectious Diseases,
Canisius Wilhelmina Hospital, Nijmegen, The Netherlands): Interlaboratory
compatibility of microsatellite based fungal typing.
12.00-12.15
Marrigje Nabuurs-Franssen, Jacques F. Meis, Ton Dofferhoff & Andreas Voss
(Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina
Hospital, Nijmegen, The Netherlands): Intravenous antimycotica use in daily clinical
practice.
12.15-12.30
Tom Sprong (Department of Medical Microbiology and Infectious Diseases, Canisius
Wilhelmina Hospital, Nijmegen, The Netherlands): Cerebrale aspergillose na een kop koe botsing.
12.30-12.45
Ferry Hagen1, Eiko E. Kuramae1, Marjan Bovers1, Dave J.C. Gerits1, Collin H.A.
Gerritzen1, Wieland Meyer2 & Teun Boekhout1 (1CBS Fungal Biodiversity Center, Yeast
Research Programme, Utrecht, The Netherlands; 2Westmead Hospital, University of
Sydney, Molecular Mycology Laboratory, Westmead, NSW, Australia): Where is the
origin of the Cryptococcus gattii Vancouver Island outbreak?
12.45-13.00
Josef Kaltseis1, Johannes Rainer1 and Sybren de Hoog2 (Institute of Microbiology,
Leopold-Franzens-University Innsbruck, Austria; 2Centraalbureau voor
Schimmelcultures, Utrecht, The Netherlands): Accumulation of Pseudallescheria and
Scedosporium in urban areas - the promotion of pathogenic fungi by humans.
ABSTRACTS
Scedosporium prolificans in Germany
K. Tintelnot1, A. Haas2, B. Graf3 & M. Seibold1
1
Robert Koch-Institut, Nordufer 20, 13353 Berlin, Germany; 2Max von Pettenkofer-Inst.,
University München, Germany; 3Institute for Medical Microbiology, Charité Berlin,
Germany
Scedosporium prolificans, a mould closely related to Pseudallescheria boydii, is a rare
but emerging pathogen due to its complete resistance to traditional antimycotic treatment.
20 patients in Germany with deep infection or long lasting colonization by S. prolificans
from 1993 to March 2007 have been reviewed retrospectively. The mortality rate in all 12
patients with invasive scedosporiosis and a severe underlying disease (AML, BMT,
SOTX, AIDS, long term treatment with corticosteroids) was 100%. Another 7 patients, 6
of them with cystic fibrosis, one with chronic sinusitis, were colonized without
development of an invasive infection. All clinical isolates and reference strains from
Belgium, Spain, U.K., Australia and U.S.A. were analyzed by sequencing of the ITS and
parts of the 18S and 28SrDNA regions and of the translation elongation factor EF1-alpha.
No variability of the ITS gene and flanking regions within the species S. prolificans was
detectable. Interestingly enough sequencing of the translation elongation factor EF1alpha revealed three different genotypes of S. prolificans, which might be useful for
further epidemiological studies. In vitro susceptibility testing confirmed a certain benefit
by the combination of voriconazole and terbinafin, nevertheless on a level being far from
optimal blood levels of the antifungals.
Biodiversity of the genus Cladophialophora, agents of human infection
H. Badali1, 2, G. S. de Hoog1, A. H. G. Gerrits van den Ende1, M .J. Najafzadeh1
1
Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands; 2Department of
Medical Mycology and Parasitology, School of Medicine, Mazandaran University of
Medical Sciences, Sari, Iran
Cladophialophora is a dematiaceous hyphomycete genus that produces branched chains
of ellipsoidal to fusiform conidia through blastic, acropetal conidiogenesis. Teleomorphs
have not been found, but are predicted to belong to the ascomycete genus Capronia.
According to rDNA SSU gene this genus phylogenetically belongs to the
Herpotrichiellaceae (Chaetothyriales) a fungal group containing a large number of
human pathogens and opportunists that causative agents of chromoblastomycosis which
is a chronic subcutaneous mycotic infection caused by pigmented or dematiaceous
saprophytic moulds ubiquitous in the environment. Taxonomy of Cladophialophora until
recently was mainly based on sequence diversity of the Internal Transcribed Spacer (ITS)
region of ribosomal DNA gene. In the present study, multilocus sequence data and
phenetic characteristics were combined to characterize the morphologically similar
Cladophialophora strains from various sources.
Methods. Sequences of the ITS region, partial Elongation Factor 1-α (EF 1-α) and ßTubulin (β-TUB) genes were analyzed for a set of 43 strains belonging to the
Cladophialophora with different primers. The program MrAIC was used to select the
substitution model. According to Akaike Information Criterion (AICc) calculation was
used to select the right model for the ratio of parameters to characters for all loci. The
congruence of gene genealogies was assessed using the Partition Homogeneity Test
(PHT) in PAUP v.4.010 based on sequences of 3 loci using only parsimony-informative
sites with 1000 replicates with random additions.
Results. A total of 3 loci of 43 strains were included in the comparison. The trees had
similar resolutions and topologies with all algorithms used. From the data obtained, it is
concluded that several novel species are to be described. In addition to the common agent
of chromoblastomycosis, C. carrionii, a second, morphologically similar pathogenic
species has been described recently, differentiated by molecular data. PHT showed that
tree gene lineages were not congruent (P = 0.02). Morphological observations between
group A and B revealed that group A generally had conidiophores that arise at right
angles from creeping hyphae while in group (B) tend to be ascending, hyphae gradually
becoming conidiophore like. Several new species isolated from the environment have
differences with these species both molecular and morphological point of view (not
shown). Recently Crous et al. (2007) have reported several new species that isolated from
the environments, which are clearly different in ITS from all species reported in this
poster.
Discussion. Concerning SSU rDNA phylogeny data, all Cladophialophora species that
are consistently associated with pathology to humans belong to the Herpotrichiellaceae
in the order Chaetothyriales. Within this order, the genus Cladophialophora is
polyphyletic. De Hoog et al. (2007) have shown that a new environmental species of
Cladophialophora, C. yegresii, was reported which occurs sympatrically in Venezuela in
the endemic region of C. carrionii. Ribosomal sequences indicated that other
environmental and plant pathogenic species are found at larger distances. The set of 43
strains contained two further new Cladophialophora species, isolated from ice tea and
alkyl benzene-polluted soil, respectively, in addition to a single strain representing an
undescribed agent of chromoblastomycosis.
Polymorphisms in genes involved in innate immunity predispose towards myetoma
susceptibility
Wendy W.J. van de Sande1, Ahmed Fahal2, Henri Verbrugh1 & Alex van Belkum1
1
Department of Bacteriology, University of Rotterdam, The Netherlands; 2Mycetoma
Research Center, University of Khartoum, Sudan
Madurella mycetomatis is the main causative agent of mycetoma, a tumorous fungal
infection characterized by infiltration of large numbers of neutrophils at the site of
infection. In endemic areas the majority of inhabitants have antibodies to M.
mycetomatis, though only a small proportion of individuals actually develop mycetomal
disease. It, therefore, appears that neutrophils are unable to clear the infection in some
individuals. To test this hypothesis, eleven single nucleotide polymorphisms (SNPs)
involved in neutrophil function were studied in a population of Sudanese mycetoma
patients versus geographically and ethnically matched controls. Significant differences in
allele distribution for interleukin 8 (CXCL8), its receptor CXCR2, thrombospondin-4
(TSP-4), nitric oxide synthase 2 (NOS2) and complement receptor 1 (CR1) were found.
Further, the NOS2 Lambaréné polymorphism was clearly associated with lesion size. The
genotypes obtained for CXCL8, its receptor CXCR2 and TSP-4 all predisposed to a
higher CXCL8 expression in patients, which was supported by the detection of
significantly elevated levels of CXCL8 in patient serum. The NOS2 genotype observed in
healthy controls was correlated with an increase in NOS2 expression and higher
concentrations of nitrate and nitrite in control serum. We present the first evidence of
human genetic predisposition towards susceptibility to mycetoma, a neglected infection
of the poor.
Molecular characterisation of azole-resistant Aspergillus fumigatus isolates
E. Snelders, H.A.L. van der Lee, J. Kuijpers, A.J.M.M. Rijs, W.J.G. Melchers & P.E.
Verweij
University Medical Center St. Radboud, Nijmegen, The Netherlands
Background: Multiple-triazole-resistance (MTR) was recently described in 9 patients
from 6 different hospitals in the Netherlands. There was a significant increase of MTR in
A. fumigatus compared with a previously performed survey (1945 to 1998), but the
prevalence of MTR is unknown. We investigated the prevalence of azole-resistance in A.
fumigatus in a prospectively collected fungus culture collection over a period of 13 years.
We characterized the isolates phenotypically and genotypically and determined the
number of cases of azole-resistant aspergillosis.
Methods: All Aspergillus isolates that were cultured from clinical specimens between
1994 and 2006 were routinely stored and included in the study. The A. fumigatus isolates
were screened for the MTR phenotype by subculturing on slants containing 8 mg/l
itraconazole. Any isolate showing growth was investigated for phenotype (MIC testing,
CLSI M38-A) and genotype (sequencing of cyp51 gene, β-tubulin gene and calmodulin
gene, and microsatellite analysis). Control patients and strains from every month between
2002 and 2006 were randomly selected and compared for phenotype and underlying
diseases.
Results: A total of 1,753 A. fumigatus isolates from 1,119 patients were screened for the
MTR phenotype. All MTR cases were observed after 1999 with an annual prevalence of
1.6% to 6%. All MTR isolates were highly resistant to itraconazole, voriconazole,
posaconazole and ravuconazole. For amphotericin B, terbinafin and caspofungin there
was no difference of MICs between MTR and non MTR isolates. Sequence analysis of
the cyp51A gene showed a substitution of leucine 98 for histidine, together with two
copies of a 34 bp sequence in tandem in the gene promoter, in 25 of 26 of itraconazoleresistant isolates. Microsatellite analysis indicated that all azole-resistant isolates were
genetically distinct but clustered.
Conclusions: Azole-resistance has emerged in A. fumigatus since the year 2000 with
resistant isolates causing invasive aspergillosis. Clinical microbiology laboratories need
to routinely determine the azole susceptibility of clinically relevant A. fumigatus isolates,
and physicians should be aware of azole resistance as a possible cause of treatment
failure. Since the reasons for emergence of MTR require is unknown further investigation
is needed.
Where is the origin of the Cryptococcus gattii Vancouver Island outbreak?
Ferry Hagen1, Eiko E. Kuramae1, Marjan Bovers1, Dave J.C. Gerits1, Collin H.A.
Gerritzen1, Wieland Meyer2 & Teun Boekhout1
1
CBS Fungal Biodiversity Center, Yeast Research Programme, Uppsalalaan 8, NL-3584
CT, Utrecht, The Netherlands; 2Westmead Hospital, University of Sydney, Molecular
Mycology Laboratory, Westmead, NSW 2145, Australia
The pathogenic basidiomycetous yeast Cryptococcus gattii may cause a life-threatening
disease of the central nervous system, lungs and skin in humans and animals. C. gattii is
found mainly in tropical and sub-tropical regions of South America, Africa, Asia and
Australia where it is endemic. Recently, a cryptococcosis outbreak in both humans and
animals occurred on Vancouver Island (British Columbia, Canada) (Kidd et al., PNAS
101, 2004). This outbreak was shown to be caused by a rare genotype of C. gattii
(AFLP6A or RAPD VGIIa). The objective of this study was to find the origin of the
outbreak isolates.
A selection of thirty-four C. gattii outbreak isolates and ninety C. gattii reference strains
were analyzed by AFLP. The six different AFLP fingerprint analyses were carried out in
duplicate. Reproducible marker fragments were used for population genetic analysis. In
addition, polymorphic fragments from the AFLP analyses were used to develop a
multilocus sequence typing (MLST) approach.
Fraser et al. (Nature 437, 2005) suggested that the Vancouver Island outbreak isolates
originated from Australia. However, our results based on AFLP and MLST analyses
show that the outbreak isolates originated from South America. South American isolates
were found to be ancestral to Australian and Asian isolates as well.
Kidd, S.E., F. Hagen, R.L. Tscharke, M. Huynh, K.H. Bartlett, M. Fyfe, L. MacDougall, T, Boekhout, K.J.
Kwon-Chung and W. Meyer: A rare genotype of Cryptococcus gattii caused the cryptococcosis outbreak
on Vancouver Island (British Columbia, Canada) (2004) Proc Natl Acad Sci USA 101:17258-17263
Fraser, J.A., S.S. Giles, E.C. Wenink, S.G. Geunes-Boyer, J.R. Wright, S. Diezmann, A. Allen, J.E. Stajich,
F.S. Dietrich, J.R. Perfect and J. Heitman: Same-sex mating and the origin of the Vancouver Island
Cryptococcus gattii outbreak (2005) Nature 437:1360-1364
Accumulation of Pseudallescheria and Scedosporium in urban areas - the promotion
of pathogenic fungi by humans
Josef Kaltseis1, Johannes Rainer1 & Sybren de Hoog2
1
Institute of Microbiology, Leopold-Franzens-University Innsbruck, Austria;
2
Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands
Members of the genera Pseudallescheria and Scedosporium (Microascaceae) are
causative agents of mycoses, among them life-threatening infections of the CNS.
Common routes of infection are the aspiration of polluted water after near-drowning
events and traumatic inoculation. Estimation of the infection risk can only be assessed
when its abundance in possibly infective materials is known. Therefore, an exact
knowledge of the habitats of Pseudallescheria and Scedosporium strains is important for
epidemiological purposes. In this study soil from three protection areas, two urban
locations and agricultural lands were sampled in Austria in the years 2004 and 2006, and
the concentration of Pseudallescheria and Scedosporium species was evaluated.
Furthermore the ammonium- and pH-values of the different soils were analysed. Isolation
was done with a semi-selective medium (SceSel+). Additionally population genetic
studies with the isolated strains were carried out. A correlation between the load of
Pseudallescheria/Scedosporium spp. and the Ammonium concentration was found. It was
proven that in soils of industrial areas, parks and playgrounds as well as in agricultural
lands the accumulation of Pseudallescheria and Scedosporium is promoted. Furthermore
all species, except S. prolificans, and molecular determined clusters were found.
Presumably those soils present a higher infection risk in case of accidents. Therefore it
should be kept in mind that Pseudallescheria/Scedosporium infections are not improbable
in case of traumatic inoculation of “urban” soil or aspiration of water with agitated
sediment.
Candida tropicalis in human disease
Louis Chai1, Peter Warn2 & David Denning2
1
University Medical Center St. Radboud, Nijmegen, The Netherlands; 2School of
Medicine, University of Manchester, U.K.
Candida tropicalis is an emerging pathogen with a global trend suggesting an increasing
incidence. The rate of invasive disease with this species vary widely by geography; and is
more common in tropical countries. It may be a nosocomial pathogen, being carried on
healthcare workers’ hands. C. tropicalis is taxonomically close to C. albicans and may
share many of the pathogenic traits although it is a rare cause of oral and vaginal mucosal
infection. Its genome has been sequenced but not analysed. It is virulent especially in
neutropenic hosts. Following candidaemia, there can be haematogenous seeding to
peripheral organs especially endophthalmitis. For candidaemia and invasive candidasis
caused by C. tropicalis, an amphotericin B preparation or echinocandin is recommended
as first-line treatment. Physicians practising in regions where C. tropicalis is common
will need to be mindful of the characteristics of this lesser-described pathogen.